Technical Field
The present disclosure relates to methods for the detection of specific nucleotide sequences and reagents and kits for use in practicing the methods.
Related Art
Nicking endonuclease enzymes have been previously described. For example, U.S. Pat. No. 6,191,267 discloses recombinant DNA encoding a nicking endonuclease, N.BstNBI, and the production of N.BstNBI restriction endonuclease from the recombinant DNA utilizing PIeI modification methylase. U.S. Pat. No. 6,395,523 discloses two methods to engineer nicking endonucleases from existing Type Ms restriction endonucleases, and the production of engineered nicking endonucleases. One approach involves inactivating the dimerization function of a Type Ms restriction enzyme using site-directed mutagenesis approach. Another approach involves replacing the cleavage domain of a Type Hs restriction enzyme with the cleavage domain from the naturally occurring nicking endonuclease, N.BstNBI.